Cov tswv yim kuaj mob ib txwm siv los kuaj xyuas cov kab mob sib kis yuav tsum tau siv cov cuab yeej benchtop uas tsis tsim nyog rau kev kuaj xyuas qhov taw qhia (POCT).Emerging microfluidics yog ib qho kev siv tshuab me me, tsis siv neeg, thiab kev sib xyaw ua ke uas yog ib txoj hauv kev zoo rau cov txheej txheem ib txwm siv rau kev ceev, tus nqi qis, qhov tseeb ntawm qhov chaw kuaj mob.Molecular diagnostic txoj kev yog dav siv nyob rau hauv cov khoom siv microfluidic li txoj kev zoo tshaj plaws rau kev kuaj kab mob.Qhov kev tshuaj xyuas no qhia txog kev nce qib tsis ntev los no hauv microfluidic-based molecular diagnostics ntawm cov kab mob sib kis los ntawm ob qho tib si kev kawm thiab kev lag luam.Ua ntej, peb piav qhia txog cov txheej txheem ntawm cov txheej txheem ntawm nucleic acids, suav nrog cov qauv ua ntej, kev nthuav dav, thiab kev nyeem cov teeb liab.Cov yam ntxwv, qhov zoo thiab qhov tsis zoo ntawm plaub hom microfluidic platforms tau muab piv rau.Tom ntej no, peb yuav sib tham txog kev siv cov kev ntsuam xyuas digital rau qhov kom muaj nuj nqis ntawm nucleic acids.Ob qho tib si classical thiab tsis ntev los no kev lag luam microfluidic-based molecular diagnostic devices tau sau tseg ua pov thawj ntawm lub xeev tam sim no ntawm kev ua lag luam.Thaum kawg, peb muab cov lus qhia yav tom ntej rau kev kuaj mob microfluidic ntawm cov kab mob sib kis.
Cov kab mob kis tau los ntawm cov kab mob, suav nrog cov kab mob, kab mob, thiab cov kab mob cab, uas tau muab faib thoob plaws ntiaj teb.Tsis zoo li lwm yam kab mob, cov kab mob kis tau sai sai thiab sib kis ntawm tib neeg thiab cov tsiaj nyeg los ntawm inoculation, huab cua thiab dej xov xwm [1].Kev tiv thaiv kab mob kis tau yog qhov tseem ceeb raws li kev ntsuas kev noj qab haus huv rau pej xeem.Peb lub tswv yim tseem ceeb rau kev tiv thaiv kab mob sib kis: (1) tswj cov kab mob;(2) cuam tshuam ntawm txoj kev sib kis;(3) tiv thaiv cov neeg raug tsim txom.Ntawm cov tswv yim tseem ceeb, kev tswj hwm qhov chaw ntawm tus kab mob yog suav tias yog lub tswv yim tseem ceeb tshaj plaws vim nws yooj yim thiab tus nqi qis.Kev kuaj mob sai, kev cais tawm, thiab kev kho mob ntawm cov neeg muaj mob yog qhov tseem ceeb, yuav tsum tau ceev ceev, rhiab, thiab raug cov tswv yim kuaj mob kom raug [2].Kev kuaj mob tam sim no feem ntau yog sib xyaw nrog kev kuaj mob raws li cov tsos mob thiab cov tsos mob thiab kev tshawb fawb hauv chaw kuaj mob xws li kab lis kev cai ntawm tes thiab kev kuaj mob molecular, uas xav tau cov neeg ua haujlwm cob qhia, cov txheej txheem ua haujlwm hnyav, thiab cov cuab yeej ntsuas kim [3, 4].Kev tiv thaiv kab mob sib kis yuav tsum tau ceev, pheej yig, thiab raug kuaj xyuas hauv zos, tshwj xeeb tshaj yog nyob rau hauv cov chaw muaj peev xwm txwv tsis pub muaj kab mob sib kis thiab mob hnyav [5], nrog rau kev kho mob hauv roob moj sab qhua lossis hauv kev sib ntaus sib tua, qhov xwm txheej kub ntxhov tsis tuaj yeem tshwm sim..kev kho mob tsuas yog txwv [6].Nyob rau hauv cov ntsiab lus no, microfluidics yog ib lub tshuab ua ke nrog microelectromechanical systems technologies, nanotechnology, los yog cov ntaub ntawv kev tshawb fawb rau cov kua dej meej manipulation [7,8,9,10], muab tshiab possibilities rau point-of-care detection (POCT).) Cov neeg kis kab mob sab nraud cov tsev kho mob thiab cov chaw kuaj mob.Piv nrog rau kev kuaj mob ib txwm siv sijhawm, microfluidic thev naus laus zis muab cov qauv thiab kev txuag nqi rau kev kuaj mob molecular thaum muaj kab mob.Kev sib kis thoob ntiaj teb ntawm tus kab mob coronavirus 2019 (COVID-19) yog tshwm sim los ntawm qhov mob hnyav ua pa nyuaj rau tus mob coronavirus 2 (SARS-CoV-2), yog li qhov tseem ceeb ntawm microfluidics rau kev tiv thaiv raws sijhawm thiab kev tswj hwm tus kabmob kis tau zoo dua [11, 12. , 13] ib.Tsis zoo li kev kuaj mob ib txwm muaj, microfluidic POCT siv cov khoom siv me me xws li cov ntsuas ntsuas ntawm lub rooj zaum mus rau me me sidestream xeem strips los ntsuas ze ntawm qhov piv txwv [14].Cov kev ntsuam xyuas no muaj cov qauv yooj yim lossis tsis muaj cov qauv npaj, kev nthuav dav lub teeb liab nrawm, thiab kev nyeem cov teeb liab uas ua rau lub sijhawm luv luv thiab cov txiaj ntsig raug nyob hauv feeb.Kev muaj thiab kev tsim khoom loj ntawm microfluidic-based cov cuab yeej kho mob tau nthuav dav lawv cov txiaj ntsig zoo thiab kev siv tshuaj ntsuam xyuas ncaj qha sab nraum tsev kho mob, nyob ze tus neeg mob, thiab txawm nyob hauv tsev.
Ntawm cov tswv yim uas twb muaj lawm los kuaj xyuas cov kab mob sib kis, kev kuaj mob molecular yog ib qho ntawm cov rhiab heev [15, 16].Tsis tas li ntawd, kev tshuaj ntsuam xyuas molecular feem ntau yog siv los ua tus qauv kub rau kev tshawb nrhiav txuas ntxiv ntawm COVID-19, tso cai rau kev tshawb pom tus kab mob tshwj xeeb ntawm thaj chaw RNA lossis DNA ua ntej qhov pib ntawm kev tiv thaiv kab mob [17, 18].Hauv kev tshuaj xyuas tam sim no, peb nthuav tawm cov kev txhim kho tshiab kawg hauv microfluidics-raws li molecular diagnostic txheej txheem rau cov kab mob sib kis, los ntawm kev kawm txog kev xav txog kev lag luam yav tom ntej (Fig. 1).Peb yuav pib nrog peb cov kauj ruam tseem ceeb hauv kev tshawb nrhiav nucleic acid: on-chip qauv pretreatment, nucleic acid amplification, thiab nyeem cov teeb liab.Peb mam li muab piv rau ntau hom microfluidic platforms nrog lawv cov qauv thiab kev ua haujlwm, uas qhia cov yam ntxwv tshwj xeeb (qhov muaj zog thiab qhov tsis muaj zog).Kev tshawb nrhiav digital nucleic acid yog sib tham ntxiv thiab muab ua piv txwv ntawm lub cim thib peb thev naus laus zis rau kev txheeb xyuas qhov tseeb ntawm cov kab mob sib kis.Tsis tas li ntawd, ob peb yam khoom siv thiab kev lag luam POCT tshiab yuav raug nthuav tawm los qhia txog lub xeev tam sim no ntawm microfluidic POCT kev lag luam rau kev kuaj mob molecular.Peb tseem yuav sib tham thiab piav qhia peb lub zeem muag rau cov ntawv thov yav tom ntej.
Modules ntawm microfluidic chips rau kev tshawb nrhiav nucleic acid tuaj yeem muab faib ua peb pawg (sampling, lees paub, thiab teeb liab) raws li lawv txoj haujlwm [19].Ntawm cov qauv no, cov qauv qauv feem ntau paub txog cov qauv lysis thiab nucleic acid extraction.Lub sensor module tsuas yog tswj kev hloov dua siab tshiab thiab nthuav dav ntawm nucleic acid signals.Lub teeb liab module pom lub teeb liab hloov dua siab tshiab thiab ua tiav los ntawm qhov ntsuas qhov ntsuas.Raws li cov txheej txheem ntawm kev kuaj pom nucleic acids ntawm ib nti, peb yuav sau cov txheej txheem ntau yam uas tuaj yeem paub txog "kev tawm tswv yim thiab kev tso tawm" muaj nuj nqi.
Thawj kauj ruam hauv kev tshawb nrhiav nucleic acid yog nucleic acid extraction, piv txwv li cais lub hom phiaj nucleic acid los ntawm cov qauv qub.Nucleic acid extraction yog ua los purify nucleic acids los ntawm lwm yam molecular contaminants, xyuas kom meej qhov kev ncaj ncees ntawm cov thawj qauv ntawm nucleic acid molecules, thiab optimize tau.Nucleic acid extraction yuav tsum muaj cov qauv tsim nyog lysis thiab nucleic acid capture, qhov zoo thiab kev ua tau zoo uas muaj kev cuam tshuam loj rau kev tshawb fawb thiab kev kuaj mob.Tej yam tsis zoo tshwm sim thaum lub sij hawm rho tawm tuaj yeem txwv kev tshawb nrhiav ntxiv.Piv txwv li, polymerase saw cov tshuaj tiv thaiv (PCR) thiab voj isothermal amplification (LAMP) txoj kev yog inhibited los ntawm ib co residual organic solvents xws li ethanol thiab isopropanol nyob rau hauv nucleic acid cais reagents [20].Kua-kua extraction thiab solid-phase extraction yog txoj kev nrov tshaj plaws rau kev cais nucleic acids [21], txawm li cas los xij, kua-kua extraction ntawm ib nti yog tsis tshua muaj kev txwv, txij li cov reagents siv hauv cov kua-kua extraction ua rau corrosion ntawm feem ntau microfluidic chips. .Ntawm no, peb qhia txog microarray-raws li cov txheej txheem rho tawm cov txheej txheem thiab sib piv lawv qhov zoo thiab qhov tsis zoo.
Silicon yog cov khoom siv substrate sib xws nrog nucleic acids vim nws biocompatibility, stability, thiab yooj yim ntawm kev hloov kho [22].Qhov tseem ceeb, thaum hloov kho nrog silica los yog lwm yam ntaub ntawv, cov khoom sib xyaw no nthuav tawm cov khoom rau adsorb tsis zoo nqi nucleic acids nyob rau hauv qis pH, siab ntsev tej yam kev mob thaum eluting nrog siab pH, tsawg ntsev daws.Raws li qhov tshwm sim no, nws muaj peev xwm ua kom huv cov nucleic acid.
Ntau hom silica-raws li cov ntaub ntawv tau siv rau nucleic acid extraction hauv microfluidics, xws li silica hlaws, hmoov, microfiber lim, thiab silica membranes [23, 24, 25, 26].Nyob ntawm cov khoom siv, cov khoom siv silicon tuaj yeem siv rau hauv microcircuits hauv ntau txoj kev.Piv txwv li, silica granules, hmoov, thiab kev lag luam nanofilters tuaj yeem tso rau hauv qhov pores lossis microchannels ntawm microfluidic chips thiab pab tshem tawm cov nucleic acids los ntawm cov qauv [27, 28, 29].Nto-hloov silica daim nyias nyias kuj tuaj yeem siv los ntxuav DNA sai sai ntawm cov kab mob ntawm tus nqi qis.Piv txwv li, Wang et al.[30] Los ntawm kev sib txuas denaturing amplification cov tshuaj tiv thaiv nrog vesicle-mediated saw exchanges nrog silica membranes coated nrog chitosan oligosaccharides, ib tug ntau yam portable system tau qhia uas ntse kuaj 102-108 colony tsim units.(CFU)/ml Vibrio parahaemolyticus., thiab lub xub ntiag ntawm tus kab mob no tau yooj yim pom.Powell et al.[31] Silicon-raws li microarrays tau siv los kuaj kab mob siab C (HCV), human immunodeficiency virus (HIV), Zika virus, thiab human papillomavirus thiab tsis siv neeg propagation, uas 1.3 μl tortuous microreactor tau tsim los ntes cov kab mob RNA.thiab ua nyob rau hauv situ amplification.Ntxiv nrog rau cov txheej txheem no, qhov chaw hloov kho silica microcolumns tseem ua lub luag haujlwm tseem ceeb hauv nucleic acid extraction, raws li cov duab geometry thiab cov khoom ntawm cov khoom hloov kho zoo heev ua rau kom muaj txiaj ntsig zoo.Chen et al.[32] tau thov kom muaj lub microfluidic platform rau kev sib cais ntawm tsawg-concentration RNA raws li amino-coated silicon microcolumns.Cov khoom siv microfluidic no sib xyaw ua ke ntawm 0.25 cm2 micropillars ntawm silicon substrate kom ua tiav cov txiaj ntsig zoo dua los ntawm thaj chaw siab rau qhov ntim piv tsim.Qhov zoo ntawm qhov tsim no yog tias cov khoom siv microfluidic tuaj yeem ua tiav txog li 95% nucleic acid extraction efficiency.Cov tswv yim silicon-based no qhia tau hais tias tus nqi ntawm kev sib cais nucleic acids sai sai ntawm tus nqi qis.Nyob rau hauv kev sib xyaw nrog microfluidic chips, silicon-raws li cov tswv yim extraction tsis tau tsuas yog ua kom cov kev ua tau zoo ntawm nucleic acid nrhiav tau, tab sis kuj pab txhawb cov miniaturization thiab kev koom ua ke ntawm cov cuab yeej tshuaj ntsuam xyuas [20].
Txoj kev sib nqus sib nqus siv cov khoom sib nqus kom cais cov nucleic acids nyob rau hauv lub xub ntiag ntawm sab nraud magnetic teb.Feem ntau siv cov khoom sib nqus muaj xws li Fe3O4 los yog γ-Fe2O3 magnetic particles coated nrog silica, amino thiab carboxyl [33,34,35,36].Qhov txawv ntawm cov khoom sib nqus piv rau silicon-based SPE txoj kev yog qhov yooj yim ntawm kev tswj thiab tswj nrog cov hlau nplaum sab nraud.
Siv cov electrostatic kev sib cuam tshuam ntawm nucleic acids thiab silica, nyob rau hauv tej yam kev mob ntawm siab ntsev thiab tsis muaj pH, nucleic acids yog adsorbed rau saum npoo ntawm silica-coated magnetic hais, thaum nyob rau hauv tej yam kev mob ntawm tsis tshua muaj ntsev thiab pH siab, molecules yuav ntxuav. dua..Silica-coated magnetic hlaws dai ua rau nws muaj peev xwm rho tawm DNA los ntawm cov qauv ntim loj (400 μL) siv cov hlau nplaum tswj [37].Raws li kev ua qauv qhia, Rodriguez-Mateos li al.[38] siv cov hlau nplaum tunable los tswj cov kev hloov ntawm cov hlaws sib nqus mus rau cov chav sib txawv.Raws li silica-coated magnetic particles, 470 daim ntawv luam / mL ntawm SARS-CoV-2 genomic RNA tuaj yeem muab rho tawm los ntawm cov qauv dej khib nyiab rau LAMP thim rov qab kuaj pom (RT-LAMP) thiab cov lus teb tuaj yeem nyeem hauv 1 teev.liab qab qhov muag (Fig. 2a).
Cov khoom siv raws li cov khoom sib nqus thiab cov khoom ntxeem tau.Conceptual diagram of the IFAST RT-LAMP microfluidic device for SARS-CoV-2 RNA detection (adapted from [38]).b Centrifugal micro ntaus ntawv rau dSPE ntawm buccal swab nucleic acid (hloov los ntawm [39]).c Built-in self-powered qauv concentrator siv daim npav FTA® (hloov los ntawm [50]).d Fusion 5 lim ntawv hloov nrog chitosan (hloov los ntawm [51]).SARS-CoV-2 mob hnyav acute respiratory syndrome coronavirus 2, RT-LAMP reverse transcription loop mediated isothermal amplification, FTA finders technology partners, NA nucleic acid
Qhov zoo them cov hlau nplaum yog qhov zoo tagnrho rau kev txuas cov phosphate qaum ntawm nucleic acid.Ntawm qee qhov ntsev concentration, qhov tsis zoo phosphate pawg ntawm nucleic acids tuaj yeem raug them zoo rau ntawm qhov chaw sib nqus sib nqus.Yog li ntawd, sib nqus nanoparticles nrog ib tug ntxhib nto thiab ib tug siab ceev ntawm cov amino pawg tau tsim rau extraction ntawm nucleic acids.Tom qab sib nqus sib cais thiab thaiv, cov hlau nplaum nanoparticles thiab DNA complexes tuaj yeem siv ncaj qha rau hauv PCR, uas tshem tawm qhov xav tau rau kev ua kom huv thiab siv sijhawm ntev thiab kev ua haujlwm elution [35].Sib nqus nanoparticles coated nrog cov pawg tsis zoo carboxyl kuj tau siv los cais cov nucleic acids adsorbed rau ntawm qhov chaw hauv siab concentration polyethylene glycol thiab sodium chloride daws [36].Nrog rau cov hlaws-hloov sib nqus sib nqus, DNA rho tawm yog sib xws nrog kev nthuav dav tom ntej.Dignan et al.[39] tau piav qhia txog lub tshuab hluav taws xob thiab nqa tau yooj yim centrifugal microfluidic platform rau nucleic acid pretreatment, tso cai rau cov neeg ua haujlwm tsis siv neeg siv nws ntawm qhov chaw.Tsis tas li ntawd, kev sib raug zoo ntawm DNA cais nrog LAMP, ib txoj hauv kev zoo haum rau kev soj ntsuam nucleic acid, ntxiv rau cov khoom siv tsawg tsawg thiab tsim nyog rau kev soj ntsuam colorimetric (Fig. 2b).
Magnetic hlaws txoj kev muaj peev xwm ntawm automated extraction, qee qhov muaj nyob rau hauv kev lag luam automated nucleic acid extractors [KingFisher;ThermoFisher (Waltham, MA, USA), QIAcube® HT;CapitalBio (Beijing, Tuam Tshoj) thiab Biomek®;Beckman (Miami, USA).), Florida, USA).Qhov zoo ntawm kev sib txuas cov hlaws sib nqus nrog cov microfluidics tuaj yeem siv rau kev ua haujlwm zoo ntawm cov nucleic acids, uas tuaj yeem ua rau kev txhim kho ntawm kev kuaj mob molecular;Txawm li cas los xij, kev sib xyaw ua ke ntawm cov hlaws sib nqus nrog microfluidics tseem vam khom ntau ntawm cov txheej txheem tswj kom meej meej ntawm cov hlaws sib nqus, uas piav qhia qhov nrov ntawm cov khoom lag luam yog loj thiab kim, uas txwv tsis pub siv cov hlaws sib nqus ntxiv hauv POCT.
Ntau cov ntaub ntawv ntxeem tau xws li hloov kho nitrocellulose lim, Finders Technology Associates (FTA) phaib, polyethersulfone-raws li cov ntaub ntawv lim, thiab cov ntaub ntawv glycan-coated kuj tau siv rau kev tshawb nrhiav nucleic acid [40, 41, 42, 43, 44].Porous fibrous cov ntaub ntawv xws li daim ntawv fibrous yog thawj zaug siv los cais DNA los ntawm lub cev entangling ntev-stranded DNA molecules nrog fibers.Cov pores me me ua rau muaj kev txwv lub cev muaj zog ntawm DNA molecules, uas cuam tshuam zoo rau DNA rho tawm.Vim muaj qhov sib txawv ntawm qhov pore loj ntawm daim ntawv fiber ntau, qhov kev ua tau zoo ntawm kev rho tawm tsis tuaj yeem ua tau raws li qhov xav tau ntawm DNA amplification [45, 46].Daim npav FTA yog daim ntawv lim dej coj mus muag siv hauv thaj chaw ntawm cov tshuaj forensic thiab dav siv hauv lwm qhov chaw ntawm kev kuaj mob molecular.Los ntawm kev siv cellulose lim ntawv impregnated nrog ntau yam tshuaj los lyse cell membranes nyob rau hauv cov qauv, lub tso tawm DNA yog tiv thaiv los ntawm degradation mus txog 2 xyoo.Tsis ntev los no, daim ntawv impregnated cellulose tau tsim los rau kev tshawb nrhiav molecular ntawm ntau yam kab mob, suav nrog SARS-CoV-2, leishmaniasis, thiab malaria [47,48,49].HIV nyob rau hauv lub plasma cais yog lysed ncaj qha, thiab cov kab mob nucleic acid yog enriched nyob rau hauv lub FTA® flow membrane ua rau hauv lub concentrator, uas tso cai rau kev ua tau zoo ntawm cov nucleic acid [50] (Fig. 2c).Qhov teeb meem tseem ceeb ntawm kev tshawb nrhiav nucleic acid siv FTA phaib yog tias cov tshuaj xws li guanidine thiab isopropanol inhibit cov tshuaj tiv thaiv tom ntej.Txhawm rau daws qhov teeb meem no, peb tsim Fusion 5 chitosan-hloov lim daim ntawv, uas sib txuas qhov zoo ntawm ob qho tib si lub cev interlacing ntawm DNA molecules thiab fiber ntau lim ntawv, thiab electrostatic adsorption ntawm DNA ntawm chitosan-hloov tebchaw kom ua tau zoo nucleic acid extraction. ..lim fibers [51] (Fig. 2d).Zoo sib xws, Zhu et al.[52] tau qhia txog chitosan-hloov PCR txoj kev raws li nyob rau hauv situ capillary microfluidic system rau kev sib cais ceev ceev thiab kuaj pom tus kab mob Zika RNA.Nucleic acids tuaj yeem adsorbed / desorbed nyob rau hauv ib qho kev sib xyaw lysate / PCR nruab nrab, feem, raws li cov khoom siv ntawm chitosan.rau thiab tawm", teb rau pH.
Raws li tau hais los saum no, cov tswv yim no muab cov txiaj ntsig zoo ntawm ntau yam khoom siv theem thiab ua kom muaj txiaj ntsig ntawm nucleic acid extraction hauv microfluidics.Hauv kev siv cov tswv yim, kev siv cov ntaub ntawv no hauv ntau qhov tsis zoo, thiab kev kho kom zoo los yog kev hloov pauv ntawm cov khoom siv nrog cov ntaub ntawv no tuaj yeem khaws lawv txoj haujlwm.Yog li ntawd, nws ntseeg tau tias qhov kev siv ntawm cov tswv yim no tom qab kev sim kev sim tuaj yeem txo cov nqi.
Nucleic acid kuaj ntawm microfluidic platforms feem ntau siv cov qauv me me (< 100 µl), yog li yuav tsum muaj kev nthuav dav ntawm lub hom phiaj nucleic acids nrog cov kev sojntsuam tshwj xeeb rau kev hloov pauv mus rau lub teeb liab uas yooj yim rau kev tshawb pom hauv qab (optical, hluav taws xob, thiab sib nqus) [53, 54] ib. Nucleic acid kuaj ntawm microfluidic platforms feem ntau siv cov qauv me me (< 100 µl), yog li yuav tsum muaj kev nthuav dav ntawm lub hom phiaj nucleic acids nrog cov kev sojntsuam tshwj xeeb rau kev hloov pauv mus rau lub teeb liab uas yooj yim rau kev tshawb pom hauv qab (optical, hluav taws xob, thiab sib nqus) [53, 54] ib. При тестировании нуклеиновых кислот на микрожидкостных платформах часто используются небольшие объемы образцов (< 100 мкл), поэтому требуется амплификация целевых нуклеиновых кислот с помощью специальных зондов для преобразования в сигнал, удобный для последующего обнаружения (оптического, электрического и магнитного) [53, 54]. Thaum kuaj cov nucleic acids ntawm microfluidic platforms, cov qauv me me (<100 µL) feem ntau yog siv, yog li kev nthuav dav ntawm lub hom phiaj nucleic acids nrog kev sojntsuam tshwj xeeb yuav tsum tau hloov nws mus rau hauv lub teeb liab yooj yim rau kev tshawb nrhiav tom qab (optical, hluav taws xob, thiab sib nqus) [53, 54] ib.微流控 上 的 核酸 核酸核酸 使用检测 使用使用使用 小样本量使用小样本量小样本量 (<100 μ因此需要 特定 特定 探针 扩增 核酸 核酸 核酸, 以 为转换 便于便于 检测核酸, 光学, 和 磁学) 的 信号) 的 信号) 的 信号) 的 信号) 的 信号) 的 信号) 的 信号) 的 信号) 的 信号) 的 信号) 的 信号) 的 信号) 的 信号) 的 信号) 的 信号). ].微流控 平台 的上 核酸的 核酸 检测使用 使用 小样本量使用小样本量 ((<100 μ需要 目标特定 目标 目标, 以以 下游目标目标, 光学以 下游下游 下游下游下游目标目标目标目标目标目标目标目标目标目标目标目标目标目标目标目标目标目标目标目标目标目标目标目标下游下游下游下游下游 (光学,, 54, 54, 54 ]. Обнаружение нуклеиновых кислот на микрожидкостных платформах обычно использует небольшие объемы образцов (<100 мкл), что требует амплификации целевых нуклеиновых кислот с помощью специальных зондов для преобразования в сигналы для последующего обнаружения (оптического, электрического и магнитного) [53, 54]]. Kev tshawb pom ntawm nucleic acids ntawm microfluidic platforms feem ntau siv cov qauv me me (<100 μl), uas yuav tsum tau ua kom muaj zog ntawm lub hom phiaj nucleic acids nrog kev sojntsuam tshwj xeeb los hloov lawv mus rau hauv cov cim rau kev tshawb pom tom qab (optical, hluav taws xob, thiab sib nqus) [53, 54]] .Nucleic acid amplification nyob rau hauv microfluidics kuj tuaj yeem ua kom muaj kev cuam tshuam, txhim kho kev kuaj pom, txo cov qauv kev xav tau, thiab txhim kho kev tshawb pom qhov tseeb [55, 56].Nyob rau hauv xyoo tas los no, nrog kev paub txog kev tshawb pom sai thiab raug, ntau txoj hauv kev nucleic acid amplification tau siv rau hauv microfluidics, suav nrog PCR thiab qee qhov kev tawm tsam isothermal amplification.Tshooj lus no yuav piav qhia txog txoj hauv kev rau kev tshawb nrhiav nucleic acid raws li microfluidic systems.
PCR yog ib qho kev simulation ntawm DNA replication txheej txheem ntawm ib tug kab mob, qhov kev tshawb xav ntawm uas tau piav nyob rau hauv kom meej nyob rau hauv lwm qhov chaw thiab yuav tsis tham txog ntawm no.PCR tuaj yeem nthuav dav me me ntawm lub hom phiaj DNA / RNA ntawm tus nqi exponential, ua PCR yog lub cuab yeej muaj zog rau kev tshawb pom sai ntawm nucleic acids.Nyob rau hauv xyoo tsis ntev los no, ntau portable microfluidic li nruab nrog PCR thermal cycling systems tau tsim kom tau raws li cov kev xav tau ntawm point-of-care diagnostics [57, 58].On-chip PCR tuaj yeem muab faib ua plaub hom (paj, nruam ntws, spatially switched, thiab convective PCR) raws li kev tswj kub sib txawv [59].Piv txwv li, Gee et al.[60] tau tsim ib txoj kev rov qab rov qab ncaj qha rau PCR (RT-qPCR) txoj hauv kev ntawm lawv tus kheej microfluidic platform rau kev tshawb nrhiav multiplex ntawm SARS-CoV-2, influenza A thiab B viruses hauv caj pas swab kuaj (Fig. 3a).Park et al.[61] tsim cov kab mob yooj yim txheeb xyuas nti los ntawm kev sib koom ua ke nyias zaj duab xis PCR, electrodes, thiab ntiv tes ua haujlwm polydimethylsiloxane-based microfluidic module.Txawm li cas los xij, ob qho tib si ua haujlwm embody qhov tsis zoo ntawm cov qauv PCR.PCR yuav tsum tau thermal cycling, uas txwv tsis pub ntxiv cov cuab yeej miniaturization thiab txo lub sijhawm sim.
Txoj kev loj hlob ntawm kev nruam ntws raws li microfluidic thiab qhov chaw hloov PCR yog qhov tseem ceeb los daws qhov teeb meem no.Siv cov serpentine channel ntev los yog luv luv ncaj channel, nruam ntws PCR tuaj yeem ua kom nrawm nrawm los ntawm kev ua kom cov reagents nyob rau hauv peb cheeb tsam preheat nrog lub twj tso kua mis tawm.Qhov kev ua haujlwm no ua tiav kom tsis txhob muaj qhov hloov pauv ntawm qhov sib txawv ntawm cov tshuaj tiv thaiv kub thiab yog li txo lub sijhawm sim [62] (Daim duab 3b).Hauv lwm txoj kev tshawb fawb los ntawm Jung et al.[63] tau npaj ib qho tshiab rotary PCR genetic analyzer uas sib txuas cov yam ntxwv ntawm kev ruaj khov thiab ntws PCR rau ultrafast thiab multiplex thim rov qab PCR (Fig. 3c).Rau nucleic acid amplification, PCR microchip yuav tig los ntawm peb qhov cua sov thaiv ntawm qhov kub sib txawv: 1. Denaturation block 94 ° C, 2. Annealing block ntawm 58 ° C, 3. Expansion block ntawm 72 ° C.
Kev siv PCR hauv microfluidics.Schematic sawv cev ntawm dirRT-qPCR ntawm lub microfluidic platform (hloov los ntawm [60]).b Schematic sawv cev ntawm lub nruam ntws PCR microarray raws li ib tug serpentine channel (hloov los ntawm [62]).c Schematic sawv cev ntawm lub teb PCR genetic analyzer, muaj xws li ib tug microchip, peb lub tshav kub thaiv thiab ib tug stepper lub cev muaj zog (hloov los ntawm [63]).d Daim duab ntawm thermoconvection PCR nrog centrifugation thiab teeb tsa (hloov los ntawm [64]).DirRT-qPCR, direct quantitative reverse transcription polymerase saw cov tshuaj tiv thaiv
Siv cov capillaries thiab loops los yog nyias daim hlau, convection PCR tuaj yeem ua kom cov nucleic acids nrawm nrawm los ntawm cov cua sov uas tsis muaj qhov xav tau rau lub twj tso kua mis sab nraud.Piv txwv li, lub cyclic olefin polymer microfluidic platform tau tsim nyob rau ntawm ib qho kev sib hloov cua sov theem uas siv thermal cycling nrog centrifugation hauv PCR voj microchannel [64] (Fig. 3d).Cov tshuaj tiv thaiv yog tsav los ntawm thermal convection, uas tsis tu ncua sib pauv siab thiab qis kub hauv microchannel nrog cov qauv sib luag.Tag nrho cov txheej txheem amplification tuaj yeem ua tiav hauv 10 feeb nrog kev kuaj pom qhov txwv ntawm 70.5 pg / channel.
Raws li qhov xav tau, PCR ceev ceev yog lub cuab yeej muaj zog rau kev sib xyaw ua ke ntawm cov qauv- teb cov qauv tshuaj ntsuam xyuas thiab kev ntsuas ntau yam.Rapid PCR txo cov sij hawm xav tau los kuaj SARS-CoV-2, uas ua rau muaj txiaj ntsig zoo rau kev tswj hwm tus kab mob COVID-19.
PCR xav tau cov thermal cycler uas tsis haum rau POCT.Tsis ntev los no, cov txheej txheem isothermal amplification tau siv rau microfluidics, suav nrog tab sis tsis txwv rau LAMP, recombinase polymerase amplification (RPA), thiab amplification raws li nucleic acid sequence [65,66,67,68].Nrog rau cov tswv yim no, nucleic acids yog amplified ntawm qhov kub thiab txias, yooj yim rau kev tsim cov nqi qis, rhiab heev portable POCT li rau kev kuaj mob molecular.
High-throughput microfluidics-raws li LAMP kev soj ntsuam tso cai rau ntau yam kev kuaj pom cov kab mob sib kis [42, 69, 70, 71].Ua ke nrog lub centrifugal microfluidic system, LAMP tuaj yeem ua kom yooj yim rau kev ua haujlwm ntawm kev tshawb nrhiav nucleic acid [69, 72, 73, 74, 75].Lub spin-and-react SlipChip yog tsim los rau qhov pom ntawm ntau cov kab mob sib luag uas siv LAMP [76] (Fig. 4a).Thaum siv lub teeb pom kev zoo hauv qhov kev ntsuam xyuas, fluorescence teeb liab-rau-lub suab sib piv yog kwv yees li 5-fold, thiab qhov kev kuaj pom tau mus txog 7.2 daim ntawv luam / μl ntawm genomic DNA. Ntxiv mus, lub hav zoov ntawm tsib kab mob plab hnyuv kab mob, xws li Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis thiab Vibrio parahaemolyticus, tau pom los ntawm txoj kev hauv <60 min. Ntxiv mus, lub hav zoov ntawm tsib kab mob plab hnyuv kab mob, xws li Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis thiab Vibrio parahaemolyticus, tau pom los ntawm txoj kev hauv <60 min.Ntxiv mus, muaj tsib yam kab mob kab mob ntawm lub plab zom mov, suav nrog Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis thiab Vibrio parahaemolyticus, tau pom siv txoj kev no tsawg dua 60 feeb.此外, 基于 在 在 在 在 在 视化 视化 视化 五 五 五 五 常见 细菌病 细菌病 细菌病细菌病 存在 存在 存在存在 存在, 大 芽孢杆菌, 大 肠杆菌, 肠 沙门 菌, 河流 弧菌 副溶血性 和 弧菌 副溶血性 菌 菌 菌菌 弧菌., 基于 在 在 在 在 在 60 五 视化 五 五 消化道 消化道 常见 细菌病常见 存在消化道 存在存在 存在, 大, 大, 肠, 弧菌肠杆菌, 弧菌弧菌 性, 弧菌弧菌 性 ... 弧菌 弧菌 弧菌弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 HIPTsis tas li ntawd, muaj tsib yam kab mob hauv plab hnyuv, suav nrog Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvius, thiab Vibrio parahaemolyticus, tau pom siv txoj kev no tsawg dua 60 feeb.
Qhov zoo ntawm LAMP hauv microfluidics suav nrog, ntawm lwm tus, cov lus teb ceev ceev thiab kev kuaj pom me me.Txawm li cas los xij, vim tias qhov kub thiab txias (nyob ib ncig ntawm 70 ° C), aerosols yog inevitably generated thaum lub sij hawm LAMP, ua rau ib tug siab tsis zoo tus nqi.Kev soj ntsuam tshwj xeeb, tsim qauv primer, thiab tswj qhov kub thiab txias kuj yuav tsum tau ua kom zoo rau LAMP.Tsis tas li ntawd, nti tsim uas siv ntau lub hom phiaj nrhiav pom ntawm ib lub nti yog qhov muaj txiaj ntsig zoo thiab yuav tsum tau tsim.Tsis tas li ntawd, LAMP yog tsim rau ntau lub hom phiaj kev tshawb nrhiav kev koom ua ke hauv ib lub nti, uas yog qhov tseem ceeb, tab sis tseem muaj ntau chav rau kev txhim kho.
Qhov siab tsis tseeb qhov zoo ntawm LAMP tuaj yeem raug txo qis nrog RPA, vim qhov ntsuas kub tsis tshua muaj tshwm sim (~ 37 ° C) ua rau muaj teeb meem evaporation tsawg [77].Hauv RPA system, ob qhov sib txawv primers pib DNA synthesis los ntawm kev khi rau recombinase thiab amplification tuaj yeem ua tiav hauv 10 feeb [78,79,80,81].Yog li ntawd, tag nrho cov txheej txheem RPA yog nrawm dua li PCR lossis LAMP.Nyob rau hauv xyoo tas los no, microfluidic thev naus laus zis tau pom tias yuav txhim kho qhov nrawm thiab raug ntawm RPA [82,83,84].Piv txwv li, Liu et al.[85] tau tsim ib qho microfluidic integrated lateral flow polymerase recombinase amplification assay rau kev tshawb nrhiav sai thiab rhiab heev ntawm SARS-CoV-2 los ntawm kev sib txuas lus rov qab RPA (RT-RPA) thiab universal lateral flow test strip detection system.nyob rau hauv ib qho microfluidic system.Daim duab 4b).Qhov txwv ntawm kev kuaj pom yog 1 daim / µl lossis 30 daim ntawv / qauv, thiab kev kuaj pom tuaj yeem ua tiav hauv li 30 feeb.Kong et al.tau tsim cov khoom siv microfluidic hnav.[86] siv lub cev kub thiab lub xov tooj ntawm tes-based fluorescence nrhiav kom tau sai thiab ncaj qha mus kuaj HIV-1 DNA siv RPA (Daim duab 4c).Qhov kev ntsuam xyuas RPA hnav tau kuaj pom 100 daim ntawv luam / mL ntawm lub hom phiaj ib ntus hauv 24 feeb, ua kom pom qhov muaj peev xwm zoo rau kev kuaj mob sai ntawm cov menyuam mos uas muaj kab mob HIV-1 hauv cov chaw muaj kev txwv.
Isothermal amplification hauv point-of-care testing (POCT).Kev tsim kho thiab tsim cov kiv thiab cov tshuaj tiv thaiv SlipChip.Tom qab plasma vuam, cov chips sab saum toj thiab hauv qab tau sib sau ua ke nrog cov txiv ntseej los ua cov nti kawg (hloov los ntawm [76]).b Schematic ntawm MI-IF-RPA system rau kev kuaj pom COVID-19 (hloov los ntawm [85]).c Schematic ntawm ib qho kev xeem RPA rau kev kuaj pom sai ntawm HIV-1 DNA (hloov los ntawm [86]).SE Salmonella enterica, VF Vibrio fluvius, VP Vibrio parahaemolyticus, BC Bacillus cereus, EC Escherichia coli, FAM carboxyfluorescein, human immunodeficiency virus HIV, RPA recombinase polymerase amplification, LED light emitting diode, MI-IF-RPA Polycomflase-RPA Amplification
Microfluidic-based RPA tab tom txhim kho sai, txawm li cas los xij, tus nqi ntawm chip fabrication thiab cov tshuaj tiv thaiv kev noj qab haus huv yog siab dhau thiab yuav tsum tau txo kom muaj cov cuab yeej siv no.Tsis tas li ntawd, lub siab rhiab heev ntawm RPA tuaj yeem cuam tshuam rau kev nthuav dav ntawm cov khoom tsis yog tshwj xeeb, tshwj xeeb tshaj yog thaum muaj kev sib kis.Cov kev txwv no tuaj yeem cuam tshuam rau kev siv RPA hauv microfluidic systems thiab ua kom zoo dua ntxiv.Cov qauv tsim zoo thiab cov kev sojntsuam rau ntau lub hom phiaj tseem xav tau los txhim kho qhov ua tau ntawm RPA-based microfluidic cov tswv yim hauv POCT.
Cas13 thiab Cas12a muaj peev xwm random cleave nucleic acids thiab yog li tsim tau raws li kev tshawb nrhiav thiab kuaj cov cuab yeej.Cas13 thiab Cas12a tau qhib ua haujlwm thaum khi rau lub hom phiaj DNA lossis RNA, feem.Thaum ua haujlwm, cov protein pib cleave lwm cov nucleic acids nyob ze, tom qab uas qhia RNAs tsom rau cov kab mob tshwj xeeb nucleic acids tuaj yeem tshem tawm cov fluorescent probes thiab tso tawm fluorescence.Raws li qhov kev xav no, Kellner et al.[87] tau tsim ib txoj hauv kev Cas13 [Specific High-sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK)], thiab Broughton li al.[88] tsim lwm txoj hauv kev raws li Cas12a [CRISPR Trans Reporter tsom DNA endonuclease (DTECR)].
Nyob rau hauv xyoo tas los no, ntau txoj hauv kev rau kev tshawb nrhiav cov nucleic acids raws li CRISPR tau tshwm sim [89, 90].Cov txheej txheem CRISPR raws li cov qauv feem ntau siv sijhawm thiab siv zog ua haujlwm vim muaj ntau yam txheej txheem suav nrog nucleic acid extraction, amplification thiab CRISPR nrhiav.Kev tso kua rau hauv huab cua tuaj yeem ua rau muaj feem cuam tshuam tsis zoo.Muab cov saum toj no, CRISPR-based systems yog qhov xav tau ntawm kev ua kom zoo dua.
Lub tshuab pneumatically tswj microfluidic platform uas tuaj yeem ua 24 kev tshuaj ntsuam nyob rau hauv tib lub sijhawm tau tsim los rau CRISPR-Cas12a thiab CRISPR-Cas13a cov ntawv thov kev tshawb nrhiav [91].Lub kaw lus tau nruab nrog lub tshuab kuaj pom fluorescence uas hla cov nucleic acid amplification thiab tuaj yeem kuaj pom femtomolar DNA thiab RNA cov qauv.Chen et al.[92] integrated recombinase amplification nrog CRISPR-Cas12a system nyob rau hauv centrifugal microfluidics (Fig. 5a).Qhov kev ua haujlwm no kov yeej qhov nyuaj ntawm kev sib koom ua ke ob txoj haujlwm no vim tias Cas12a tuaj yeem zom cov tub txib DNA thiab cuam tshuam cov txheej txheem amplification.Tsis tas li ntawd, Chen et al.[92] Tsis tas li ntawd ua ntej khaws cov tshuaj tiv thaiv cov tshuaj tiv thaiv hauv centrifugal microfluidic tswj kom ua tiav tag nrho cov txheej txheem.Hauv lwm txoj haujlwm, Silva li al.[93] tau tsim ib txoj kev kuaj mob yam tsis muaj CRISPR/Cas12a amplification thiab lub xov tooj ntawm tes los ntes SARS-CoV-2 (Fig. 5b).Qhov kev ntsuam xyuas no, lub npe hu ua lub xov tooj ntawm tes-raws li kev nthuav dav-dawb system, suav nrog CRISPR / Cas-dependent enzyme uas yog raws li smartphone pom ntawm catalase-tsim npuas cov cim hauv microfluidic raws.Kev kuaj pom muaj txiaj ntsig tsawg dua 50 daim ntawv luam / µl ntawm nucleic acid yam tsis muaj kev nthuav dav ua ntej, tag nrho cov txheej txheem los ntawm cov qauv txhaj tshuaj rau kev nyeem ntawv yuav siv sijhawm 71 feeb xwb.
Nucleic acid nrhiav txoj hauv kev raws li CRISPR.Centrifugal POCT rau kev kuaj mob sib xyaw ua ke raws li CRISPR (hloov los ntawm [92]).b Kev Tsim Kho ntawm CASCADE test rau smartphone-based analysis of SARS-CoV-2 (adapted from [93]).RAA recombinase amplification, PAM nyob ib sab protospacer motif, CRISPR clustered luv palindromic rov ua dua ntawm lub sijhawm tsis tu ncua, CASCADE system tsis muaj xov tooj ntawm tes amplification nrog CRISPR / CAS-nyob ntawm cov enzymes, 1-ethyl-3- [3-dimethylaminopropyl]carbodiimide hydrochloride EDC
Raws li cov kauj ruam kawg hauv kev tshawb nrhiav nucleic acid, qhov teeb meem nrhiav pom ncaj qha cuam tshuam txog kev kuaj mob thiab yog qhov tseem ceeb hauv kev txhim kho cov txiaj ntsig zoo, rhiab, thiab raug POCT.Cov teeb liab tuaj yeem nyeem tau siv ntau txoj hauv kev xws li fluorescent, electrochemical, colorimetric thiab cov tswv yim sib nqus.Hauv seem no, peb piav qhia txog qhov laj thawj rau txhua txoj hauv kev thiab sib piv cov kev kuaj mob molecular ntawm cov kab mob sib kis hauv microfluidics.
Fluorescence-raws li cov tswv yim tau dav siv rau POCT kev kuaj mob ntawm cov kab mob sib kis vim lawv qhov zoo tshaj plaws ntawm kev nkag siab zoo, tus nqi qis, kev ua haujlwm yooj yim, thiab kev tsom xam ntawm kev saib xyuas [94, 95].Cov tswv yim no siv cov ntawv sau fluorophores xws li fluorescent dyes thiab nanomaterials los tsim cov teeb liab pom tau (fluorescence li lossis quenching).Qhov kev tshawb pom no qhia tias cov tswv yim fluorescence tuaj yeem muab faib ua fluorescent labeling ncaj qha, teeb liab-on, thiab teeb liab-tawm fluorescent nrhiav pom [96].Direct fluorescent label detection siv cov ntawv tshwj xeeb fluorescent los sau cov ligands uas tsim cov fluorescence tshwj xeeb thaum xaiv khi rau lub hom phiaj.Rau cov teeb liab-raws li fluorescence nrhiav pom, qhov zoo ntawm lub teeb liab fluorescent yog qhov zoo rau qhov loj ntawm kev txaus siab.Fluorescence siv yog negligible nyob rau hauv tsis muaj lub hom phiaj thiab yog detectable thaum muaj ib tug txaus npaum li cas ntawm lub hom phiaj.Hloov pauv, qhov kev siv ntawm fluorescence pom los ntawm "lub teeb liab-tawm" fluorescence inversely proportional rau tus nqi ntawm lub hom phiaj, pib ncav cuag tus nqi siab tshaj plaws thiab maj mam txo qis thaum lub hom phiaj loj.Piv txwv li, siv CRISPR-Cas13a lub hom phiaj-dependent trans-cleavage mechanism, Tian li al.[97] tau tsim ib lub tswv yim tshiab los paub txog RNAs uas hla kev rov qab rov qab ncaj qha (Fig. 6a).Raws li kev khi rau cov hom phiaj sib luag RNAs, CRISPR-Cas13-RNA complex tuaj yeem qhib tau, ua rau muaj kev cuam tshuam los ntawm cov neeg sau xov xwm tsis tshwj xeeb RNAs.Lub fluorescently sau ntawv xov xwm [fluorophore (F)] yog quenched los ntawm lub quencher (Q) zoo thiab fluoresces thaum cleaved los ntawm lub tshuab txais complex.
Qhov kom zoo dua ntawm kev tshawb nrhiav electrochemical yog kev tshawb nrhiav siab, kev tsim khoom yooj yim, tus nqi qis, yooj yim nqa thiab tswj tsis siv neeg.Nws yog ib txoj hauv kev zoo tshaj plaws rau kev siv POCT.Raws li graphene teb-zoo transistors Gao li al.[98] tsim ib lub nanobiosensor rau multiplex nrhiav tau ntawm Lyme kab mob antigens los ntawm Borrelia burgdorferi cov kab mob nrog rau kev kuaj pom txwv ntawm 2 pg / mL (Fig. 6b).
Colorimetric kev soj ntsuam tau siv hauv POCT daim ntawv thov, tau txais txiaj ntsig los ntawm qhov zoo ntawm kev txav mus los, tus nqi qis, kev npaj tau yooj yim, thiab kev nyeem pom.Kev kuaj pom xim tuaj yeem siv cov oxidation ntawm peroxidase los yog peroxidase zoo li nanomaterials, kev sib sau ntawm nanomaterials, thiab qhov sib ntxiv ntawm cov xim dyes los hloov cov ntaub ntawv hais txog qhov muaj lub hom phiaj nucleic acids rau pom kev hloov xim [99, 100, 101].Qhov tseem ceeb, cov nanoparticles kub tau siv dav hauv kev txhim kho cov tswv yim colorimetric, thiab vim lawv muaj peev xwm ua kom muaj kev hloov xim sai thiab tseem ceeb, muaj kev txaus siab ntxiv rau kev txhim kho POCT colorimetric platforms rau hauv qhov chaw kuaj mob ntawm cov kab mob sib kis [102].Nrog cov khoom siv centrifugal microfluidic sib xyaw ua ke [103], cov kab mob hauv cov zaub mov hauv cov qauv mis tsis huv tuaj yeem kuaj pom ntawm qib 10 cov kab mob, thiab cov txiaj ntsig tuaj yeem nyeem pom hauv 65 feeb (Fig. 6c).
Cov tswv yim sib nqus sensing tuaj yeem txheeb xyuas cov tshuaj ntsuam xyuas siv cov khoom sib nqus, thiab tau muaj kev txaus siab rau POCT daim ntawv thov nyob rau xyoo tsis ntev los no.Magnetic sensing cov tswv yim muaj qee qhov tshwj xeeb zoo xws li cov khoom siv hlau nplaum uas tsis muaj nqi ntau dua li cov khoom siv kho qhov muag kim.Txawm li cas los xij, kev siv lub teb sib nqus ua kom zoo dua qhov kev tshawb pom thiab txo cov sij hawm ua piv txwv [104].Tsis tas li ntawd, cov txiaj ntsig ntawm kev soj ntsuam magnetic qhia tau hais tias muaj qhov tshwj xeeb, rhiab heev, thiab siab teeb liab-rau-suab nrov piv vim qhov tsis tseem ceeb sib nqus tom qab teeb liab ntawm cov qauv lom neeg [105].Sharma et al.integrated ib magnetic tunnel junction raws biosensor rau hauv lub portable microchip platform.[106] rau multiplex nrhiav pom cov kab mob (Fig. 6d).Biosensors rhiab heev ntes subnanomolar nucleic acids cais los ntawm cov kab mob.
Hom teeb liab nrhiav tau txoj kev.Lub tswv yim ntawm hyperlocalized nrhiav pom ntawm Cas13a (hloov los ntawm [97]).b Graphene nanobiosensor FET ua ke nrog Lyme GroES scFv (hloov los ntawm [98]).c Colorimetric indications rau multiplex nrhiav pom cov zaub mov muaj kab mob nyob rau hauv ib tug centrifugal microfluidic nti: No. 1 thiab No. 3 cov qauv nrog cov hom phiaj pathogens, thiab No. .d Biosensor raws li kev sib nqus qhov sib nqus, suav nrog lub platform, lub tshuab ua kom muaj zog, lub zog tswj, thiab lub zog hluav taws xob rau cov teeb liab tsim / tau txais (kho los ntawm [106]).GFET Graphene FET, Escherichia coli, Escherichia coli, Salmonella typhimurium, Vibrio parahaemolyticus, Vibrio parahaemolyticus, Listeria monocytogenes, PC PC, PDMS Dimethicone, PMMA polymethyl methacrylate
Txawm hais tias cov yam ntxwv zoo heev ntawm cov txheej txheem tshawb nrhiav saum toj no, lawv tseem muaj qhov tsis zoo.Cov txheej txheem no tau muab piv (table 1), suav nrog qee cov ntawv thov nrog cov ntsiab lus (pros thiab cons).
Nrog rau kev txhim kho ntawm microfluidics, microelectromechanical systems, nanotechnology thiab cov ntaub ntawv kev tshawb fawb, kev siv cov microfluidic chips rau kev tshawb nrhiav cov kab mob sib kis tau nce mus ntxiv [55,96,107,108].Kev tswj xyuas meej ntawm cov khoom siv me me thiab cov kua dej ua rau muaj kev kuaj pom tseeb thiab kev siv nyiaj zoo.Yog li ntawd, txhawm rau txhim kho ntxiv, kev siv zog tau ua kom zoo dua qub thiab txhim kho cov chips, ua rau muaj ntau yam microfluidic chips nrog cov qauv sib txawv thiab kev ua haujlwm.Ntawm no peb luv luv qhia ntau hom microfluidic platforms thiab sib piv lawv cov yam ntxwv (pros thiab cons).Tsis tas li ntawd, feem ntau ntawm cov piv txwv hauv qab no tsom ntsoov rau kev sib ntaus sib tua SARS-CoV-2.
LOCCs yog cov feem ntau miniaturized complex analytical systems thiab lawv cov kev khiav hauj lwm yog heev miniaturized, integrated, automated thiab parallelized los ntawm cov qauv txhaj tshuaj thiab kev npaj, khiav tswj thiab ua kua [109, 110].Cov kua dej yog siv los ntawm kev ua tib zoo tsim cov duab geometry thiab kev sib cuam tshuam ntawm ntau lub cev muaj zog xws li siab gradients, capillary action, electrodynamics, magnetic fields thiab acoustic waves [111].LOCC qhia tau hais tias muaj txiaj ntsig zoo nyob rau hauv kev soj ntsuam siab thiab ntau yam kev tshawb nrhiav, nrog rau kev soj ntsuam ceev ceev, cov qauv me me, kev siv hluav taws xob tsawg, thiab kev tswj xyuas thiab kev ua haujlwm siab;Txawm li cas los xij, LOCC cov cuab yeej siv tau zoo heev, thiab kev tsim khoom, ntim khoom, thiab kev sib tshuam.Txawm li cas los xij, multiplexing thiab rov siv dua ntsib teeb meem loj [96].Piv nrog rau lwm lub platform, LOCC muaj qhov tshwj xeeb zoo ntawm cov ntawv thov ntau haiv neeg thiab kev siv thev naus laus zis zoo tshaj plaws, tab sis nws qhov tsis zoo kuj tseem pom tseeb, uas yog qhov nyuaj siab thiab tsis zoo rov ua dua.Kev vam khom rau cov twj sab nraud, uas feem ntau loj thiab kim, ntxiv txwv lawv siv hauv POCT.
Thaum lub sijhawm COVID-19 tshwm sim, LOCC tau txais kev saib xyuas ntau.Nyob rau tib lub sijhawm, muaj ob peb lub chips tshiab uas ua ke nrog ntau lub thev naus laus zis.Piv txwv li, cov xov tooj smartphones tam sim no tau siv dav los ua cov khoom siv tshuaj ntsuam xyuas thiab muaj peev xwm zoo rau LOCC kev koom ua ke.Sun et al.[21] fabricated ib microfluidic nti uas tso cai multiplexing tej nucleic acid sequences ntawm tsib kab mob, xws li SARS-CoV-2, siv LAMP thiab soj ntsuam lawv siv lub smartphone nyob rau hauv 1 teev tom qab qhov kawg ntawm cov tshuaj tiv thaiv.Raws li lwm qhov piv txwv, Sundah et al.[112] tsim ib qho kev hloov pauv molecular [catalytic amplification los ntawm molecular hloov pauv xeev hloov (CATCH)] rau kev kuaj pom ncaj qha thiab nkag siab ntawm SARS-CoV-2 RNA lub hom phiaj siv smartphones. CATCH yog sib xws nrog portable LOCC thiab ua tiav kev ua tau zoo (kwv yees li 8 RNA luam / μl; < 1 h ntawm chav tsev kub) [112]. CATCH yog sib xws nrog portable LOCC thiab ua tiav kev ua tau zoo (kwv yees li 8 RNA luam / μl; < 1 h ntawm chav tsev kub) [112]. CATCH совместим с портативным LOCC и обеспечивает превосходную производительность (примерно 8 Кнопечивает превосходную производительность (примерно 8 Кноприй/1 Рприй) CATCH yog sib xws nrog portable LOCC thiab muab kev xa tawm zoo heev (kwv yees li 8 RNA luam / µl; < 1 h ntawm chav tsev kub) [112]. CATCH 与便携式LOCC 兼内并具有卓越的性能（大约8 RNA 拷贝/μl；室温下< 1 小时）[112]. CATCH 与便携式LOCC 兼内并具有卓越的性能（大约8 RNA 拷贝/μl；室温下< 1 小时）[112]. CATCH совместим с портативными LOCC и обладает превосходной производительностью (примерно 8 чрадает превосходной производительностью (примерно 8 копий) < 1 сратий Ратий CATCH yog sib xws nrog portable LOCCs thiab muaj kev ua tau zoo heev (kwv yees li 8 RNA luam / µl; < 1 teev ntawm chav tsev kub) [112].Tsis tas li ntawd, LOCC cov cuab yeej rau kev kuaj mob molecular kuj siv qee qhov kev tsav tsheb xws li lub tshuab nqus tsev, ncab, thiab hluav taws xob.Kang et al.[113] ua qauv qhia lub sijhawm tiag tiag, nrawm nrawm nanoplasma-on-a-chip PCR rau kev kuaj mob sai thiab ntau yam ntawm COVID-19 hauv thaj teb uas siv lub tshuab nqus tsev plasmonic kua PCR nti.Li et al.[114] tom qab ntawd tau tsim ib lub zog-tsav microfluidic nti uas ua rau muaj kev kuaj mob ntawm COVID-19.Lub platform siv RT-LAMP amplification system los txiav txim seb tus qauv puas zoo lossis tsis zoo.Tom qab ntawd, Ramachandran et al.[115] ua tiav qhov tsim nyog hluav taws xob gradients siv isotachophoresis (ITP), ib qho kev xaiv ion tsom rau cov txheej txheem siv hauv microfluidics.Nrog ITP, lub hom phiaj RNA los ntawm cov qauv nasopharyngeal swab tuaj yeem ua kom huv si.Tom qab ntawd Ramachandran et al.[115] Ua ke no ITP purification nrog ITP-enhanced LAMP thiab CRISPR kev soj ntsuam kuaj pom SARS-CoV-2 nyob rau hauv tib neeg nasopharyngeal swab thiab soj ntsuam kuaj hauv li 35 feeb.Tsis tas li ntawd, cov tswv yim tshiab tau tshwm sim tas li.Jadhav et al.[116] tau npaj ib txoj kev kuaj mob raws li saum npoo-txhim kho Raman spectroscopy ua ke nrog cov khoom siv microfluidic uas muaj cov vertically oriented kub / nyiaj-coated carbon nanotubes lossis pov tseg electrospun micro / nanotubes.Membrane-functionalized built-in lim microchannels yog pov tseg.Cov cuab yeej adsorbs cov kab mob los ntawm ntau lub cev kua / exudations xws li qaub ncaug, nasopharynx thiab kua muag.Yog li, tus kab mob titer muaj ntau thiab tus kab mob tuaj yeem raug txheeb xyuas los ntawm Raman kos npe.
LOAD yog lub centrifugal microfluidic platform nyob rau hauv uas tag nrho cov txheej txheem yog tswj los ntawm ib tug zaus raws tu qauv uas rotates ib tug microstructured substrate [110].Lub LOAD ntaus ntawv yog tus cwj pwm los ntawm kev siv lub zog centrifugal ua ib qho tseem ceeb tsav tsheb.Cov kua kuj raug rau capillary, Euler thiab Coriolis rog.Siv cov cuab yeej centrifuge, kev tshuaj xyuas tau ua tiav hauv kev ua haujlwm ua kua txuas ntxiv los ntawm lub radial sab hauv mus rau sab nraud, tshem tawm qhov xav tau ntxiv rau cov tubing sab nraud, twj tso kua mis, actuators, thiab active li qub.Hauv luv luv, ib txoj kev tswj hwm yooj yim ua haujlwm.Lub zog ua haujlwm ntawm cov kua hauv tib lub microfluidic channel ntawm tib qhov kev ncua deb ntawm qhov chaw thauj khoom sib npaug, uas ua rau nws tuaj yeem rov ua cov qauv channel.Yog li, LOAD cov cuab yeej siv tau yooj yim thiab pheej yig dua rau kev tsim thiab tsim khoom dua li cov khoom siv LOCC, thaum cov kev cuam tshuam yog ywj siab thiab sib luag;Txawm li cas los xij, vim muaj lub zog txhua yam ntawm cov khoom siv centrifugal, cov khoom siv nti muaj tsawg thiab cov ntim me me yog qhov nyuaj.mus rau lub tsheb.Nyob rau tib lub sijhawm, cov khoom siv LOAD feem ntau yog tsim los rau kev siv nkaus xwb, uas yog kim heev rau kev kuaj pom loj [96, 117, 118, 119].
Nyob rau hauv xyoo tsis ntev los no, LOAD, suav hais tias yog ib qho ntawm cov khoom siv zoo tshaj plaws microfluidic, tau txais kev saib xyuas zoo los ntawm cov kws tshawb fawb thiab cov tuam txhab tsim khoom.Yog li, LOAD tau txais kev lees paub dav dav thiab tau siv rau kev kuaj mob molecular ntawm cov kab mob sib kis [120, 121, 122, 123, 124], tshwj xeeb tshaj yog thaum muaj kev sib kis COVID-19.Piv txwv li, thaum kawg ntawm 2020, Ji et al.[60] tau ua pov thawj ncaj qha RT-qPCR kev soj ntsuam rau kev kuaj pom sai thiab ua haujlwm sib luag ntawm SARS-CoV-2 thiab kab mob khaub thuas A thiab B hauv caj pas swab.Then Xiong et al.[74] nthuav tawm LAMP-integrated discoid microfluidic platform rau ceev, raug, thiab ib txhij nrhiav pom ntawm xya tus tib neeg ua pa tus mob coronaviruses, suav nrog SARS-CoV-2, hauv 40 feeb.Thaum ntxov 2021, de Oliveira li al.[73] tau qhia txog polystyrene toner centrifugal microfluidic nti, manually ua nrog tus ntiv tes taw rotator, rau RT-LAMP molecular kuaj mob ntawm COVID-19.Tom qab ntawd, Dignan et al.[39] nthuav tawm lub tshuab hluav taws xob portable centrifuge microdevice rau kev ua kom huv ntawm SARS-CoV-2 RNA ncaj qha los ntawm buccal swab seem.Medved et al.[53] tau thov ib qho inline SARS-CoV-2 aerosol sampling system nrog lub ntim me me tig microfluidic fluorescent nti nrog kev kuaj pom tsis pub dhau 10 daim ntawv luam / μL thiab lub voj voog yam tsawg kawg nkaus ntawm 15 feeb.Suarez et al.[75] tsis ntev los no tau tshaj tawm txog kev txhim kho ntawm kev sib koom ua ke ntawm modular centrifugal microfluidic platform rau kev tshawb nrhiav ncaj qha ntawm SARS-CoV-2 RNA hauv cov qauv siv hluav taws xob tsis siv neeg nasopharyngeal swab siv LAMP.Cov piv txwv no qhia txog cov txiaj ntsig zoo thiab kev cog lus ntawm LOAD hauv kev kuaj mob molecular ntawm COVID-19.
Nyob rau hauv 1945 Muller thiab Clegg [125] thawj zaug nthuav tawm microfluidic raws ntawm daim ntawv siv lim ntawv thiab paraffin.Xyoo 2007, Whitesides pawg [126] tau tsim thawj daim ntawv ua haujlwm rau kev kuaj cov protein thiab piam thaj.Daim ntawv tau dhau los ua qhov zoo tshaj plaws substrate rau microfluidics.Cov ntawv muaj cov khoom siv xws li hydrophilicity thiab ntxeem tau qauv, zoo heev biocompatibility, lub teeb yuag, yooj, foldability, tsis tshua muaj nqi, yooj yim siv thiab yooj yim.Classical µPADs muaj cov qauv hydrophilic/hydrophobic ua rau ntawm daim ntawv substrates.Nyob ntawm cov qauv peb-dimensional, μPADs tuaj yeem muab faib ua ob-dimensional (2D) thiab peb-dimensional (3D) μPADs.2D µPADs yog tsim los ntawm kev tsim hydrophobic ciam teb los tsim microfluidic raws, thaum 3D µPADs feem ntau yog tsim los ntawm cov txheej txheej ntawm 2D microfluidic daim ntawv, qee zaum los ntawm daim ntawv folding, plam txoj kev, qhib channel, thiab 3D luam ntawv [96].Aqueous lossis biological kua ntawm μPAD feem ntau yog tswj los ntawm capillary quab yuam yam tsis muaj lub zog sab nraud, ua kom yooj yim khaws cia ua ntej ntawm cov tshuaj reagents, coj mus kuaj, thiab kuaj pom ntau yam.Txawm li cas los xij, kev tswj xyuas qhov tseeb thiab kev txheeb xyuas qhov sib npaug yog cuam tshuam los ntawm kev tshawb pom tsis txaus, kev nkag siab, thiab rov siv tau [96, 127, 128, 129, 130].
Raws li qhov txawv txav microfluidic platform, μPAD tau nthuav dav thiab tsim kho rau kev kuaj mob molecular ntawm cov kab mob sib kis xws li HCV, HIV, thiab SARS-CoV-2 [131, 132].Rau kev xaiv thiab rhiab nrhiav pom HCV, Tengam li al.[133] tau tsim ib qho tshiab biosensor raws li daim ntawv fluorescent siv cov tshuaj nucleic acid tshwj xeeb raws li pyrrolidinyl peptide.Nucleic acids yog covalently immobilized ntawm ib nrab oxidized cellulose daim ntawv los ntawm reductive alkylation ntawm amino pawg thiab aldehyde pawg, thiab nrhiav kom tau raws li fluorescence.Cov teeb liab no tuaj yeem nyeem tau los ntawm cov khoom siv tshwj xeeb uas muaj lub koob yees duab fluorescent portable nrog rau lub koob yees duab ntawm tes.Tom qab ntawd, Lu et al.[134] tsim ib daim ntawv-raws li hloov tau electrode raws li nickel/kub nanoparticles/carbon nanotubes/polyvinyl cawv organometallic moj khaum composites rau HIV lub hom phiaj nrhiav pom los ntawm DNA hybridization siv methylene xiav raws li ib tug DNA redox qhia.Tsis ntev los no, Chowdury et al.[135] tau nthuav tawm lub tswv yim tsim qauv tsim rau qhov taw qhia ntawm kev saib xyuas µPAD kev sim siv cov qaub ncaug cov neeg mob nyoos ua ke nrog LAMP thiab cov cuab yeej siv tau yooj yim rau kev kuaj xyuas COVID-19.
Lateral flow tests coj cov kua dej los ntawm capillary rog thiab tswj cov dej txav los ntawm kev ntub dej thiab cov yam ntxwv ntawm cov ntxeem tau lossis microstructured substrates.Cov khoom siv sab nraud muaj xws li cov qauv, conjugate, incubator thiab kuaj pom, thiab cov ntaub nqus dej.Cov nucleic acid molecules hauv LFA paub txog cov binders tshwj xeeb uas tau khaws cia ua ntej ntawm qhov chaw sib khi thiab khi ua ke.Raws li cov kua dhau los ntawm cov incubation thiab nrhiav kom pom daim hlau, cov complexes raug ntes los ntawm cov capture molecules nyob rau ntawm qhov kev xeem thiab tswj kab, uas qhia tau hais tias yuav tau nyeem ncaj qha rau ntawm qhov muag liab qab.Feem ntau, LFA tuaj yeem ua tiav hauv 2-15 feeb, uas sai dua li kev tshawb pom ib txwm muaj.Vim lub tshuab tshwj xeeb, LFA yuav tsum tau ua haujlwm tsawg thiab tsis xav tau cov cuab yeej siv ntxiv, uas ua rau nws siv tau zoo heev.Nws yog ib qho yooj yim los tsim thiab miniaturize, thiab tus nqi ntawm cov ntawv-raws li substrates yog qis dua.Txawm li cas los xij, nws tsuas yog siv rau kev soj ntsuam zoo xwb, thiab kev tshawb nrhiav kom muaj nuj nqis yog qhov nyuaj heev, thiab lub peev xwm multiplexing thiab throughput muaj tsawg heev, thiab tsuas yog ib qho txaus nucleic acid tuaj yeem kuaj pom ib zaug [96,110,127].
Txawm hais tias feem ntau cov ntawv thov ntawm LFA tau tsom mus rau cov tshuaj tiv thaiv kab mob, kev siv LFA rau kev kuaj mob molecular hauv microfluidic chips kuj ua tau zoo thiab nrov [136].In case of hepatitis B virus, HIV and SARS-CoV-2 LFA Gong et al.[137] tau npaj siab hloov pauv nanoparticle LFA platform thiab ua kom pom qhov ua tau zoo ntawm lub platform me me thiab nqa tau yooj yim los ntawm kev nkag siab thiab ntsuas ntau ntawm ntau lub hom phiaj xws li HBV nucleic acid.Tsis tas li ntawd, Fu et al.[138] ua qauv qhia LFA tshiab raws li qhov ua tau zoo ntawm Raman spectroscopy rau kev txheeb xyuas ntau ntawm HIV-1 DNA ntawm qhov tsis tshua muaj siab.Rau kev tshawb pom sai thiab rhiab ntawm SARS-CoV-2, Liu et al.[85] tsim microfluidic-integrated RPA lateral flow analysis los ntawm kev sib txuas RT-RPA thiab universal lateral flow detection system rau hauv ib qho microfluidic system.
Daim ntawv thov ntawm ntau lub microfluidic platforms sib txawv nyob ntawm seb cov kev tshawb fawb tshwj xeeb, siv tag nrho cov peev txheej thiab qhov zoo ntawm cov platforms.Nrog cov nqi pheej yig, cov twj tso kua mis thiab cov kav dej, LOCC yog lub platform zoo tshaj plaws rau kev siv ntau haiv neeg thiab kev sib koom tes nrog chav loj tshaj plaws rau kev txhim kho.Yog li ntawd, peb cia siab thiab pom zoo tias cov kev tshawb fawb tshiab tshaj plaws yuav tsum tau ua ntawm LOCC ua thawj zaug thiab kom cov xwm txheej tau ua kom zoo.Tsis tas li ntawd, cov txheej txheem ua tau zoo thiab raug yuav tsum tau tshawb pom thiab siv hauv qhov system.LOAD ua tau zoo hauv kev tswj xyuas cov kua dej los ntawm cov khoom siv LOCC uas twb muaj lawm thiab ua kom pom qhov zoo ntawm ib qho kev tsav los ntawm lub zog centrifugal yam tsis tas yuav muaj cov tsav sab nraud, thaum cov lus teb sib npaug tuaj yeem sib cais thiab sib dhos.Yog li, yav tom ntej, LOAD yuav dhau los ua lub hauv paus microfluidic platform nrog kev ua haujlwm tsawg dua thiab cov thev naus laus zis ntau dua thiab siv tau.Lub µPAD platform sib txuas cov txiaj ntsig ntawm LOCC thiab cov ntaub ntawv raws li tus nqi qis, kev siv ib leeg.Yog li ntawd, kev txhim kho yav tom ntej yuav tsum tsom mus rau cov cuab yeej yooj yim thiab tsim tau zoo.Tsis tas li ntawd, LFA yog qhov tsim nyog rau kev kuaj pom qhov muag liab qab, cog lus tias yuav txo qis kev noj zaub mov thiab ua kom nrawm dua.Cov ncauj lus kom ntxaws platform sib piv yog qhia hauv Table 2.
Cov kev soj ntsuam digital faib cov qauv mus rau ntau lub microreactors, txhua tus muaj cov lej sib cais ntawm cov hom phiaj molecules [139, 140].Cov kev ntsuam xyuas digital muab cov txiaj ntsig tseem ceeb rau kev ua tiav qhov ntau los ntawm kev ua ntau txhiab qhov kev sim biochemical tib lub sijhawm thiab tus kheej hauv micron scale compartments es tsis yog nyob rau theem txuas ntxiv.Piv nrog rau cov tshuaj microfluidics, cov tshuaj tiv thaiv kab mob tuaj yeem txo qis cov qauv ntim, nce cov tshuaj tiv thaiv kev ua haujlwm, thiab tau yooj yim sib xyaw nrog lwm cov kev tshuaj ntsuam xyuas yam tsis tas yuav muaj cov channel, twj, li qub, thiab cov qauv tsim [141, 142, 143, 144, 145, 146, 147] ib.Ob txoj hauv kev hauv qab no yog siv rau hauv cov kev ntsuam xyuas digital kom ua tiav cov kev sib cais thiab kev sib cais ntawm cov kev daws teeb meem, suav nrog cov tshuaj reagents thiab cov qauv xws li cov hlwb, nucleic acids, thiab lwm cov khoom los yog cov molecules: (1) poob emulsions siv cov kua interface tsis ruaj khov;(2) array faib yog ua los ntawm geometric txwv ntawm cov cuab yeej.Hauv thawj txoj kev, cov tee uas muaj cov tshuaj reagents thiab cov qauv hauv microchannels tuaj yeem tsim los ntawm cov txheej txheem passive xws li co-tam sim no, crossflow, ndlwg focusing, staged emulsification, microchannel emulsification, thiab membranes los ntawm viscous shear rog thiab emulsification nrog channel hloov.localization [143, 145, 146, 148, 149] los yog siv txoj kev nquag [150, 151], uas qhia ntxiv zog los ntawm hluav taws xob, sib nqus, thermal thiab mechanical control.Nyob rau hauv txoj kev tom kawg, qhov zoo tshaj plaws ntawm cov dej ntim uniformity nyob rau hauv microfluidic chambers yog sib koom los ntawm kev ua kom spatial qauv ntawm tib qhov loj, xws li micropits thiab nto arrays [152,153,154].Qhov tseem ceeb, cov tee dej yog cov dej ntws loj uas tseem tuaj yeem tsim thiab tswj tau ntawm electrode arrays raws li digital microfluidics (DMF).Electrowetting ntawm dielectrics yog ib qho ntawm qhov zoo tshaj plaws kawm DMF txoj kev xav, txij li electrowetting ntawm dielectrics tso cai rau kev tswj hwm tus neeg tee, tswj cov duab ntawm cov kua thiab cov hluav taws xob asymmetric dhau los ntawm ntau sab [141, 144].Cov haujlwm tseem ceeb nrog cov tee dej hauv DMF suav nrog kev txheeb xyuas, sib cais, thiab kev sib koom ua ke [151, 155, 156], uas tuaj yeem siv rau hauv ntau qhov kev tshuaj xyuas, tshwj xeeb tshaj yog hauv kev tshawb nrhiav molecular [157, 158, 159].
Kev kuaj pom cov nucleic acid yog qhov thib peb tiam molecular diagnostic thev naus laus zis tom qab cov qauv PCR thiab ntau npaum li cas ntawm PCR (qPCR), nyob rau tib lub sijhawm nrog kev sib tw siab thiab ua kua biopsy.Hauv ob xyoos dhau los, cov nucleic acids digital tau tsim sai heev hauv kev tshawb nrhiav molecular ntawm cov kab mob sib kis [160, 161, 162].Kev txheeb xyuas qhov tseeb ntawm cov nucleic acid kuaj pom pib nrog ntim cov qauv thiab cov tshuaj reagents rau hauv ib tus neeg cov khoom kom ntseeg tau tias txhua lub hom phiaj muaj qhov tshwm sim zoo ib yam ntawm kev nkag mus rau txhua tus neeg.Raws li txoj cai, txhua ntu yuav raug muab ntau lub hom phiaj ua ntu zus, lossis yuav tsis muaj kev ywj pheej microreaction.Los ntawm ntau yam kev hnov lus tau piav qhia saum toj no, cov khoom sib txuas nrog microbial lub hom phiaj ua ntu zus uas tsim cov teeb liab saum toj ib qho chaw pib tuaj yeem pom nrog lub qhov muag liab qab lossis los ntawm lub tshuab thiab tau sau tias qhov zoo, thaum lwm qhov chaw uas tsim cov teeb liab hauv qab qhov pib raug sau tias zoo. .cov tsis zoo, uas ua rau lub teeb liab rau txhua ntu ib boolean.Yog li, los ntawm kev suav cov naj npawb ntawm cov khoom tsim thiab tus nqi ntawm cov txiaj ntsig zoo tom qab cov tshuaj tiv thaiv, cov ntawv luam thawj ntawm cov qauv kuaj tuaj yeem sib tw siv cov qauv kev faib tawm Poisson yam tsis tas yuav tsum muaj tus qauv nkhaus, uas yuav tsum tau ua kom muaj nuj nqis niaj hnub soj ntsuam xws li. ua qpcr.[163] Piv nrog rau cov txheej txheem kuaj molecular ib txwm, kev kuaj pom cov nucleic acid muaj qib siab dua ntawm automation, siab dua kev soj ntsuam ceev thiab rhiab heev, tsawg dua reagents, tsis muaj paug, thiab yooj yim tsim thiab tsim.Rau cov laj thawj no, kev siv cov kev ntsuam xyuas digital, tshwj xeeb tshaj yog cov kev poob qis, rau kev kuaj mob molecular, sib txuas cov kev nthuav dav thiab cov teeb liab nyeem cov tswv yim, tau kawm tau zoo thaum muaj qhov tshwm sim tseem ceeb ntawm SARS-CoV-2.Piv txwv li, Yin et al.[164] ua ke cov tshuaj yaj yeeb digital thiab ceev PCR txoj hauv kev txhawm rau kuaj ORF1ab, N, thiab RNase P cov noob hauv SARS-CoV-2 hauv microfluidic nti.Qhov tseem ceeb, lub kaw lus muaj peev xwm txheeb xyuas qhov teeb meem zoo li ntawm 115 vib nas this, uas yog nrawm dua li cov PCR cov qauv, qhia txog nws cov txiaj ntsig hauv qhov chaw saib xyuas (Daim duab 7a).Dong et al.[165], Tso et al.[157], Chen YJ et al.[166] and Alteri et al.[167] kuj tau siv droplet digital PCR (ddPCR) txhawm rau kuaj SARS-CoV-2 hauv microfluidic system nrog cov txiaj ntsig zoo.Txhawm rau txhim kho qhov kev kuaj pom, Shen li al.[168] ua tiav ddPCR-raws li nti duab nyob rau hauv tsawg li 15 s yam tsis tau siv cov duab stitching cov tswv yim, ua kom ceev cov txheej txheem ddPCR thev naus laus zis los ntawm kev sim mus rau daim ntawv thov.Tsis tsuas yog thermal amplification txoj kev xws li PCR yog siv, tab sis kuj isothermal amplification txoj kev yog siv los ua kom yooj yim cov tshuaj tiv thaiv kab mob thiab teb ceev ceev.Lu et al.[71] tsim SlipChip rau kev soj ntsuam cov tee dej, muaj peev xwm tsim cov tee dej ntawm ntau qhov ntau thiab tsawg ntawm qhov siab ntom ntom hauv ib kauj ruam thiab ntsuas SARS-CoV-2 nucleic acids siv cov LAMP (Daim duab 7b).Raws li cov thev naus laus zis hloov pauv sai, CRISPR tuaj yeem ua lub luag haujlwm tseem ceeb hauv kev tshawb nrhiav cov nucleic acid los ntawm kev kos duab yooj yim colorimetric yam tsis tas yuav muaj cov nucleic acid ntxiv.Ackerman et al.tsim cov tshuaj tiv thaiv combinatorial matrix rau multiplex kev ntsuam xyuas ntawm nucleic acids.[158] kuaj pom 169 tus kab mob cuam tshuam rau tib neeg, suav nrog SARS-CoV-2, hauv cov tee uas muaj CRISPR-Cas13-raws li nucleic acid kuaj pom cov tshuaj reagents hauv microwell assay (Daim duab 7c).Tsis tas li ntawd, isothermal amplification thiab CRISPR thev naus laus zis tuaj yeem siv rau hauv tib lub zog los ua ke cov txiaj ntsig ntawm ob qho tib si.Park et al.[169] Ib qho CRISPR/Cas12a digital assay tau tsim nyob rau hauv kev lag luam microfluidic nti rau kev tshawb pom ntawm kev rho tawm thiab tua hluav taws kub-tua SARS-CoV-2 raws li ib-theem RT-RPA nrog luv thiab siab dua teeb liab-rau-rov qab nrhiav pom. lub sij hawm piv., wider dynamic range thiab rhiab heev (Fig. 7d).Qee cov lus piav qhia ntawm cov piv txwv no tau muab rau hauv Table 3.
Hom digital platform rau kev tshawb nrhiav nucleic acid.a Kev ua haujlwm ceev ceev ntawm PCR muaj plaub kauj ruam tseem ceeb: kev npaj ua qauv, kev faib tawm ntawm cov tshuaj tiv thaiv sib xyaw, cov txheej txheem nthuav dav, thiab lub hom phiaj kom muaj nuj nqis (hloov los ntawm [164]).b Schematic qhia kev tsom xam ntawm SlipChip droplets rau kev tsim cov tee dej ntawm qhov ntom ntom (hloov los ntawm [71]).c CARMEN-Cas workflow diagram13 (hloov los ntawm [158]).d Txheej txheem cej luam ntawm kev kuaj pom tus kab mob siab tshaj plaws nrog CRISPR/Cas hauv ib lub lauj kaub (hloov los ntawm [169]).W/O dej-hauv-roj, polydimethylsiloxane PDMS, PCR polymerase chain reaction, DAQ data collection, PID proportional integral derivative, CARMEN combinatorial matrix reaction for multiplex nucleic acid evaluation, SARS-CoV-2, heavy acute respiratory syndrome, coronavirus 2, RT Amplification ntawm thim rov qab transcriptase recombinase polymerase-RPA, S / B teeb liab hauv keeb kwm yav dhau
Post lub sij hawm: Sep-15-2022
